Methods of culturing and expanding mesenchymal stem cells
A method of culturing mesenchymal stem cells (MSCs) is provided. The method comprising culturing a population of the MSCs in a medium comprising an aryl hydrocarbon receptor antagonist, thereby culturing MSCs.
1. A method of culturing mesenchymal stem cells (MSCs) comprising culturing a population of the MSCs in a medium comprising an aryl hydrocarbon receptor antagonist, thereby culturing MSCs, wherein said aryl hydrocarbon receptor inhibitor comprises 4-(2-(2-(benzo[b]thiophen-3-yl)-9-isopropyl-9H-purin-6-ylamino) ethyl) phenol (StemReginin 1, SR1).
2. A method of expanding a population of mesenchymal stem cells, the method comprising culturing a seeded population of mesenchymal stem cells for a period of time sufficient for cell expansion, wherein said culturing is in a medium comprising an aryl hydrocarbon receptor antagonist, thereby generating an expanded population of mesenchymal stem cells, and wherein said aryl hydrocarbon receptor inhibitor comprises 4-(2-(2-(benzo[b]thiophen-3-yl)-9-isopropyl-9H-purin-6-ylamino) ethyl) phenol (StemReginin 1, SR1).
3. A method of generating cells useful for transplantation into a subject, the method comprising:
(a) culturing mesenchymal stem cells according to the method of claim 1 to generate a population of cultured mesenchymal stem cells;
(b) contacting said population of cultured mesenchymal stem cells with a differentiating agent, thereby generating cells useful for transplantation into a subject.
4. A method of generating cells useful for transplantation, the method comprising:
(a) expanding mesenchymal stem cells according to the method of claim 2 ; and
(b) contacting the mesenchymal stem cells with a differentiating agent, thereby generating cells useful for transplantation.
5. A cell culture, comprising mesenchymal stem cells and a medium which comprises an aryl hydrocarbon receptor antagonist, wherein said aryl hydrocarbon receptor inhibitor comprises 4-(2-(2-(benzo[b]thiophen-3-yl)-9-isopropyl-9H-purin-6-ylamino) ethyl) phenol (StemReginin 1, SR1).
6. The cell culture of claim 5 , wherein said medium further comprises at least one of nicotinamide and a growth factor.
7. The method of claim 1 , wherein the mesenchymal stem cells are derived from a tissue selected from the group consisting of bone marrow, adipose tissue, placenta and umbilical cord blood.
8. The method of claim 1 , wherein the concentration of said SR1 in said medium is at a range of 100-1000 nM.
9. The method of claim 2 , wherein the concentration of said SR1 in said medium is at a range of 100-1000 nM.
10. The method of claim 1 , wherein said medium further comprises nicotinamide at a range of 1-20 mM.
11. The method of claim 10 , wherein said medium further comprises FGF-4.
12. The method of claim 1 , wherein a calcium concentration of said medium is greater than 1.8 mM.
13. The method of claim 1 , wherein said culturing is effected on a plastic surface.
14. The method of claim 1 , wherein said medium is devoid of platelet derived growth factor (PDGF).
15. The method of claim 2 , wherein said expanding is effected under conditions that do not induce differentiation of said mesenchymal stem cells.
16. The method of claim 2 , wherein said seeded population of mesenchymal stem cells were seeded in an absence of nicotinamide.
17. The method of claim 2 , wherein said seeded population of mesenchymal stem cells were seeded in a presence of nicotinamide.
18. The method of claim 3 , wherein said differentiation agent comprises a growth factor.
19. The cell culture of claim 5 , wherein the concentration of said SR1 in said medium is at a range of 100-1000 nM.